Small volume PCR in PDMS biochips with integrated fluid control and vapour barrier

In this paper we demonstrate a new method for microfabricating,a PDMS devices that controls vapour diffusion, thereby reducing water loss at elevated temperatures and greatly increasing the reliability of the PCR. In the past, the vapour and liquid diffusion properties of the PDMS material in microfluidic devices have impaired performance. We show that this water loss is primarily due to vapour diffusion from the PDMS biochip and by implanting a polyethylene vapour barrier layer in the PDMS. the overall fluid loss was almost eliminated (reduced by a factor of 3). We have also developed a procedure to ensure irreversible bonding between the PDMS and the implant. With this improved microfabrication method we demonstrate the feasibility and advantages of performing, mall Volume PCR genetic amplification (i.e. with less than 2 mu l of PCR sample) within a PDMS-glass hybrid biochip. Diaphragm pumps and pinch-off valves were integrated in the system and these enabled fluid retention during the amplification stage and will facilitate higher levels of on-chip automation, (c) 2005 Elsevier B.V. All rights reserved.